Background: Urinary tract infection (UTI) is a kind of disease in kidneys that often caused by Escherichia coli bacteria, especially in women. Objectives: We studied the antibacterial effects of ZnO nanoparticles On patients with urinary tract infection that isolated from Alzahra hospital in Isfahan, Iran. It is hoped that by the research community's attention to this great problem of multi drug resistance E. coli treatment by ZnO NPs. Methods: In this experimental study, urine samples of multi drug resistance E. coli bacteria, Also colloidal ZnO nanoparticles prepared in size 20 nm, spherical shape and a concentration of 200 ppm were used .The methods of research were wells and disk agar diffusion in nutriant agar & LB medium and was evaluated antibacterial properties of the nanoparticles ,then incubated for 72 hours and the diameter of inhibitory zone of growth was measured, The data were analyzed by spss 16 version software. Results: The highest resistance of E. coli to antibiotics belongs to cefazolin and cefexime. The results showed that ZnO NPs in size 20 nm and spherical shape after 72 hours at 37°C can be used to treat Urinary infections caused by E. coli, it is recommended that further research considered the effects of different infections of Urinary tract infection in In vitro condition. Conclusions: Using ZnO nanoparticles with 20 nm diameters have acceptable antibacterial effect on Escherichia coli isolated from Urinary tract infection. The current study suggests that ZnO NPs could be a good candidate for an effective antibacterial agent against E. coli.